Abstract 4768: PARP inhibitor-induced autophagy provides an adaptive mechanism of drug resistance in preclinical models of ovarian cancer

Abstract

Abstract Poly (ADP) ribose polymerase inhibitors (PARPi) have shown promising activity against recurrent ovarian cancers, particularly in women with germ line mutations of BRCA1/2. The efficacy of PARP inhibitors can, however, be decreased by acquired drug resistance. Autophagy or “self-eating” is one mechanism implicated in cancer progression and therapy resistance. Autophagy is a catabolic mechanism that can degrade organelles and long-lived proteins to provide energy for cancer cells under nutrient poor conditions or in the presence of stress. The energy provided from autophagy is used to fuel the DNA repair process and metabolic needs in cancer cells. Autophagy can protect cancer cells from chemotherapy or can enhance the response to certain drugs. In this study, we have asked whether autophagy contributes to the resistance of ovarian cancer cells to PARPi. Our group has found that four PARP inhibitors (olaparib, niraparib, rucaparib and talazorapib) induce autophagy in 8 ovarian cancer cell lines, detected by LC3A to LC3B conversion on western blot analysis and punctate GFP-LC3 fluorescence. Transmission electron microscopy confirmed the increased autophagosome formation in olaparib treated cells. To test whether autophagy contributes to the resistance of OC cells to olaparib, we determined the impact of autophagy inhibition on the sensitivity of OC cell lines to olaparib by either using chloroquine, hydroxychloroquine or LYS05 to block hydrolysis of proteins and lipids in autophagosomes or using siRNA against ATG5 or ATG7 to prevent formation of autophagosome. Enhancement of PARPi activity was observed in 6 out 8 ovarian cancer cell lines. Moreover, a combination of olaparib and chloroquine proved more effective than either single agent in a xenograft model of the OVCAR8 ovarian cancer cell line and in a patient derived xenograft model. Mechanistically, olaparib decreased PARP activity leading to DNA damage and accumulation of γ-H2AX. Olaparib treatment increased reactive oxygen species (ROS) and phosphorylation of ATM, while decreasing the phosphorylation of AKT and mTOR. We also observed an increase in PTEN, a well-recognized negative regulator of AKT. Inhibition of PTEN decreases LC3 conversion, suggesting PTEN as an important regulator of olaparib-induced autophagy in ovarian cancer cells. Taken together, our data demonstrate for the first time that olaparib induces autophagy in ovarian cancer cells and provides an adaptive mechanism of resistance to PARPi. Olaparib-induced autophagy may depend upon activation of ATM and PTEN by ROS, downregulating pAKT/pmTOR signaling. Moreover we have identify unique combinations of therapy with PARPi and CQ or other inhibitors of autophagy that could improve outcomes for ovarian cancer patients. Citation Format: Janice M. Santiago-O'Farrill, Saravut J. Weroha, Xiaonan Hou, Lan Pang, Philip Rask, Zhen Lu, Robert C. Bast. PARP inhibitor-induced autophagy provides an adaptive mechanism of drug resistance in preclinical models of ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4768.